Genetic Diversity in Colletotrichum gloeosporioides from Stylosanthes spp. at Centers of Origin and Utilization

ABSTRACT Using molecular markers, this work compares the genetic diversity in Colletotrichum gloeosporioides infecting species of the tropical forage legume Stylosanthes at the center of origin in Brazil and Colombia with that of Australia, China, and India, where Stylosanthes spp. have been introduced for commercial use. There was extensive diversity in the pathogen population from Brazil, Colombia, China, and India. The Australian pathogen population was least diverse probably due to its geographical isolation and effective quarantine. The extensive diversity in China and India means that threats from exotic pathogen races to Stylosanthes pastures can potentially come from countries outside the South American center of origin. In Brazil and India, both with native Stylosanthes populations, a high level of genetic differentiation in the pathogen population was associated with sites where native or naturalized host population was widely distributed. There was limited genetic diversity at germplasm evaluation sites, with a large proportion of isolates having identical haplotypes. This contrasts recent pathogenicity results for 78 of the Brazilian isolates that show hot spots of complex races are more common around research stations where host germplasm are tested, but few are found at sites containing wild host populations. For a pathogen in which the same races arise convergently from different genetic backgrounds, this study highlights the importance of using both virulence and selectively neutral markers to understand pathogen population structure.     

Understanding patterns of tree adoption on farms in semi-arid and sub-humid Ethiopia

Trees on farms are a widespread feature of landscapes across a large part of Ethiopia with an important role in enhancing the resilience of smallholder livelihoods through the provision of ecosystem services. Despite their importance, little is known about what trees are planted or retained from natural regeneration by different types of farmers that results in the pattern of tree cover found in the region. We address this knowledge gap through analysis of household survey data from semi-arid and sub humid areas of Oromia regional state. A set of composite variables that represent distinctive patterns of tree cover on farms were derived from principal component analysis and Pearson correlation analysis. This revealed two major tree adoption strategies: farmer managed natural regeneration (FMNR) of trees to meet subsistence needs as well as contributing to other ecosystem services; and, high value agroforestry (HVAF) involving planted trees used largely to produce fruits, timber and fodder. Regression analysis further identified fine-scale variation in ecological and socio-economic factors that affect which of these two broad strategies are adopted by farmers. Favorable climatic conditions coupled with institutional arrangements to control free grazing were pre-conditions for HVAF, whereas poor biophysical potential and sloping land provided a positive incentive for farmers to adopt FMNR. Farmers with preferences for tree species with multiple utilities and locational flexibility favored FMNR while adoption of HVAF was more asset-driven. Our findings reveal that farmers integrate many native and exotic tree species on their farms to meet their variable farm conditions, needs and asset profiles in stark contrast to most tree promotion efforts that focus on a few, usually exotic, tree species. We recommend that future agroforestry promotion should embrace a diversity of tree species appropriate to matching the fine scale variation in ecological conditions and farmer circumstances encountered in the field.     

臂形草内生菌特异DNA片段的克隆及其分子鉴定

进行了臂形草内生菌特异DNA片段的克隆及分子鉴定方法的研究。从5个臂形草品种分离的5个内生菌纯培养特中，提取基因组DNA，通过140条随机引物的RAPD分析，引物OPAK10扩增出1条约500bp的共有PCR谱带。此DNA片段命名为BE1。对BE1进行分离、纯化，经点杂交证实BE1为臂形草内生菌特异DNA片段。进一步将BE1片段进行回收、克隆和DNA序列分析。BE1在基因库中进行序列分析，未发现相关同源片段。臂形草内生菌特异DNA片段的获得，为建立一种准确、快速检测臂形草内生菌及特异内生菌方法奠定了分子生物学基础。     

Genetic Diversity in South American Colletotrichum gloeosporioides Isolates from Stylosanthes guianensis, a Tropical Forage Legume

The degree of genetic diversity of 127 Colletotrichum gloeosporioides isolates from Stylosanthes guianensis genotypes in South America was measured at the molecular level by random amplified polymorphic DNA (RAPD) with nine arbitrary primers of 10 bases, and by restriction fragment length polymorphism (RFLP) with a non-LTR (long terminal repeats) retrotransposon DNA sequence. The RAPD products revealed scorable polymorphism among the isolates, and a total of 80 band positions were scored. Sixty-three of the 127 isolates were clustered into 13 distinct lineages usually correlating with geographic origin. Where isolates from various regions were clustered together, most had identical host genotype origin. The pathogen population sampled from Carimagua, Colombia, a long-time Stylosanthes breeding and selection site, with a savanna ecosystem, was highly diverse. A set of 12 S. guianensis genotype differentials was used to characterize pathogenic variability of 104 isolates and their virulence patterns were grouped into 57 pathotypes. However, when they were tested on four Australian differentials, they grouped into 11 pathotypes. As shown in previous studies, no strict correlations existed between genetic diversity measured by RAPD or RFLP, and pathotype defined by pathogenicity pattern on the differentials. Southern blot analysis of the 127 isolates revealed 23 hybridizing fragments, resulting in 41 fingerprint patterns among the 127 isolates. Relationships between RFLP and RAPD variables were examined using Spearman's Rank Correlation Coefficient, which showed that the two measures of genotypic variation are in agreement.     

Weather-based prediction of anthracnose severity using artificial neural network models

Data were collected and analysed from seven field sites in Australia, Brazil and Colombia on weather conditions and the severity of anthracnose disease of the tropical pasture legume Stylosanthes scabra caused by Colletotrichum gloeosporioides . Disease severity and weather data were analysed using artificial neural network (ANN) models developed using data from some or all field sites in Australia and/or South America to predict severity at other sites. Three series of models were developed using different weather summaries. Of these, ANN models with weather for the day of disease assessment and the previous 24 h period had the highest prediction success, and models trained on data from all sites within one continent correctly predicted disease severity in the other continent on more than 75% of days; the overall prediction error was 21·9% for the Australian and 22·1% for the South American model. Of the six cross-continent ANN models trained on pooled data for five sites from two continents to predict severity for the remaining sixth site, the model developed without data from Planaltina in Brazil was the most accurate, with >85% prediction success, and the model without Carimagua in Colombia was the least accurate, with only 54% success. In common with multiple regression models, moisture-related variables such as rain, leaf surface wetness and variables that influence moisture availability such as radiation and wind on the day of disease severity assessment or the day before assessment were the most important weather variables in all ANN models. A set of weights from the ANN models was used to calculate the overall risk of anthracnose for the various sites. Sites with high and low anthracnose risk are present in both continents, and weather conditions at centres of diversity in Brazil and Colombia do not appear to be more conducive than conditions in Australia to serious anthracnose development.     

几丁质酶基因在大肠杆菌中的表达及其酶对柱花草胶孢炭疽病原菌的抑菌效果

构建了几丁质酶基因植物表达载体,该基因在大肠杆菌中能表达35kD的蛋白质。几丁质酶粗提物在体外对柱花草胶孢炭疽病原菌有极显著的抑菌效果。